Expression of apxIA of Actinobacillus pleuropneumoniae in Saccharomyces cerevisiae

Shin, Sung Jae; Bae, Jong Lye; Cho, Young Wook; Yang, Moon Sik; Kim, Dae Hyuk; Jang, Yong Suk; Yoo, Han Sang

doi:10.4142/jvs.2003.4.3.225

Cited by 9 publications

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“…Here, we examined the efficacy of the oral administration of S. cerevisiae expressing apx IIA cloned from a A. pleuropneumoniae Korean isolate, because the gene is common in A. pleuropneumoniae serotypes 2 and 5, which are the most prevalent serotypes in Korea [24,25]. The identity of the gene was confirmed through sequence comparison with that reported in GenBank.…”

Section: Discussionmentioning

confidence: 99%

“…The cloned gene was completely sequenced with an automatic sequencer (ABI PRSIM 377XL) after analysis with restriction enzymes and expressed in E. coil M15 with pQE31 (Qiagen) via IPTG induction. Expressed protein was analyzed by SDS–PAGE and Western blot using a monoclonal antibody against recombinant ApxIIA (rApxIIA) as described previously [24,25]. rApxIIA was then purified with Ni–NTA resin (Qiagen Co.) following the manufacture's description and used for subsequent experiments.…”

Section: Methodsmentioning

confidence: 99%

“…To clone in S. cerevisiae with the YEpGPD expression vector, appropriate restriction enzyme sites were generated by subcloning apx IIA gene in the pBluescript II KS cloning vector into Escherichia coli Top 10 and the yeast expression vector was transformed into the expression host S. cerevisiae 2805 using the LiAc method [25,26]. Transformed colonies were cultured onto a selective medium (URA − ; yeast nitrogen base 6.7 g, casamino acid 5 g, glucose 20 g, adenine 0.03 g, tryptopan 0.03 g, and bactoagar 20 g in 1000 ml of DW) for 12 h, transferred into basic medium (YEPD; yeast extract 10 g, bactopeptone 20 g, and glucose 20 g in 1000 ml of DW) and cultured for 2–3 days at 30 °C until 0.6–0.7 at OD 600 .…”

Section: Methodsmentioning

confidence: 99%

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Induction of antigen-specific immune responses by oral vaccination withSaccharomyces cerevisiaeexpressingActinobacillus pleuropneumoniaeApxIIA

Shin

Bae

Cho

et al. 2005

FEMS Immunology &Amp; Medical Microbiology

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An effective way of inducing both mucosal and systemic immune responses to protect against Actinobacillus pleuropneumoniae serotype 2 Korean isolate was examined in mice by oral immunization using Saccharomyces cerevisiae expressing the ApxIIA protein. The immunogenicity of the yeast-derived ApxIIA antigen was confirmed by the challenge test and ApxIIA-specific IgG antibody response assay. The group subcutaneously immunized with the protein extracted from the yeast expressing ApxIIA showed a higher survival rate after challenging with A. pleuropneumoniae serotype 2 isolate and IgG antibody level in serum than the group injected with that prepared from the yeast harboring vector only. Feeding the yeast expressing ApxIIA to mice induced both systemic and mucosal immune responses against the antigen. ApxIIA-specific IgA antibody titers and the number of IgA-secreting cells of mice vaccinated with S. cerevisiae expressing ApxIIA dose-dependently increased from the third immunization in both intestine and lung (P<0.01). A similar tendency of ApxIIA-specific IgG antibody responses was observed in the sera. The protective efficacy of the oral immunization was then evaluated by a challenge with a minimal lethal dose (MLD, 4.5 x 10(7) CFU/ml) of the A. pleuropneumoniae serotype 2 isolate. Fifty percent of the 30 mg administered group and 30% of the 15 mg administered group survived while none of the mice in the control groups survived after 36 h. These results suggest that feeding animals the yeast expressing the antigen can be an effective strategy to induce protective immune responses against A. pleuropneumoniae infection.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Induction of antigen-specific immune responses by oral vaccination withSaccharomyces cerevisiaeexpressingActinobacillus pleuropneumoniaeApxIIA

Shin

Bae

Cho

et al. 2005

FEMS Immunology &Amp; Medical Microbiology

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“…pleuropneumoniae serovar 5. The sequences obtained corresponding to the apxIA gene of the isolates were divided into 14 haplotypes and represented 3066 bp, corresponding to the complete sequence of the apxIA gene [56].…”

Section: Resultsmentioning

confidence: 99%

Polymorphism analysis of the apxIA gene of Actinobacillus pleuropneumoniae serovar 5 isolated in swine herds from Brazil

et al. 2018

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The bacterium Actinobacillus pleuropneumoniae is the etiological agent of Contagious Porcine Pleuropneumonia, a disease responsible for economic losses in the swine industry worldwide. A. pleuropneumoniae is capable of producing proteinaceous exotoxins responsible for inducing hemorrhagic lesions, one of which is ApxI. Few studies have conducted an in-depth evaluation of polymorphisms of the nucleotides that make up the ApxI toxin gene. Here we analyze the polymorphisms of the apxIA gene region of A. pleuropneumoniae serovar 5 isolated from swine in different regions in Brazil and report the results of molecular sequencing and phylogenetic analysis. Analysis of the apxIA gene in 60 isolates revealed the presence of genetic diversity and variability. The polymorphisms in the nucleotide sequences determined the grouping of the Brazilian sequences and five more sequences from the GenBank database into 14 different haplotypes, which formed three main groups and revealed the presence of mutations in the nucleotide sequences. The estimation of selection pressures suggests the occurrence of genetic variations by positive selective pressure on A. pleuropneumoniae in large groups of animals in relatively small spaces. These conditions presumably favor the horizontal dissemination of apxIA gene mutations within bacterial populations with host reservoirs. As a result, the same serovar can demonstrate different antigenic capacities due to mutations in the apxIA gene. These alterations in sequences of the apxIA gene could occur in other areas of countries with intense swine production, which could lead to differences in the pathogenicity and immunogenicity of each serovar and have implications for the clinical status or diagnosis of A. pleuropneumoniae.

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“…For the oral vaccine, S. cerevisiae expressing ApxIA or ApxIIA antigens were prepared as previously described [34,35]. …”

Section: Methodsmentioning

confidence: 99%

Enhancement of protective immune responses by oral vaccination with Saccharomyces cerevisiae expressing recombinant Actinobacillus pleuropneumoniae ApxIA or ApxIIA in mice

Shin

Kang

et al. 2007

J Vet Sci

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We previously induced protective immune response by oral immunization with yeast expressing the ApxIIA antigen. The ApxI antigen is also an important factor in the protection against Actinobacillus pleuropneumoniae serotype 5 infection; therefore, the protective immunity in mice following oral immunization with Saccharomyces cerevisiae expressing either ApxIA (group C) or ApxIIA (group D) alone or both (group E) was compared with that in two control groups (group A and B). The immunogenicity of the rApxIA antigen derived from the yeast was confirmed by a high survival rate and an ApxIA-specific IgG antibody response (p < 0.01). The highest systemic (IgG) and local (IgA) humoral immune responses to ApxIA and ApxIIA were detected in group E after the third immunization (p < 0.05). The levels of IL-1β and IL-6 after challenge with an A. pleuropneumoniae field isolate did not change significantly in the vaccinated groups. The level of TNF-α increased in a time-dependent manner in group E but was not significantly different after the challenge. After the challenge, the mice in group E had a significantly lower infectious burden and a higher level of protection than the mice in the other groups (p < 0.05). The survival rate in each group was closely correlated to the immune response and histopathological observations in the lung following the challenge. These results suggested that immunity to the ApxIA antigen is required for optimal protection.

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Expression of apxIA of Actinobacillus pleuropneumoniae in Saccharomyces cerevisiae

Cited by 9 publications

References 0 publications

Induction of antigen-specific immune responses by oral vaccination withSaccharomyces cerevisiaeexpressingActinobacillus pleuropneumoniaeApxIIA

Induction of antigen-specific immune responses by oral vaccination withSaccharomyces cerevisiaeexpressingActinobacillus pleuropneumoniaeApxIIA

Polymorphism analysis of the apxIA gene of Actinobacillus pleuropneumoniae serovar 5 isolated in swine herds from Brazil

Enhancement of protective immune responses by oral vaccination with Saccharomyces cerevisiae expressing recombinant Actinobacillus pleuropneumoniae ApxIA or ApxIIA in mice

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